Overview

  • Product nameAnti-ENPP2 antibody [5H3]
    See all ENPP2 primary antibodies
  • Description
    Mouse monoclonal [5H3] to ENPP2
  • Tested applicationsSuitable for: IHC-P, ELISA, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Recombinant full length protein of Human ENPP2.

  • Positive control
    • In Western Blot, this antibody gave a positive signal in HEK293 whole cell lysate and in the following human tissue lysates: kidney; placenta; ovary; small intestine. In IHC, this antibody gave a positive signal in human tonsil paraffin sections. In IF/ICC, this antibody gave a positive signal in HeLa cells.
  • General notes

    Alternative versions available:

    Anti-ENPP2 antibody (Alexa Fluor® 647) [5H3] (ab196712)

Properties

Applications

Our Abpromise guarantee covers the use of ab77104 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-P Use a concentration of 1 µg/ml.
ELISA Use at an assay dependent concentration.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 100 kDa (predicted molecular weight: 99 kDa).
ICC/IF Use at an assay dependent concentration.

Target

  • FunctionHydrolyzes lysophospholipids to produce lysophosphatidic acid (LPA) in extracellular fluids. Major substrate is lysophosphatidylcholine. Also can act on sphingosylphosphphorylcholine producing sphingosine-1-phosphate, a modulator of cell motility. Can hydrolyze, in vitro, bis-pNPP, to some extent pNP-TMP, and barely ATP. Involved in several motility-related processes such as angiogenesis and neurite outgrowth. Acts as an angiogenic factor by stimulating migration of smooth muscle cells and microtubule formation. Stimulates migration of melanoma cells, probably via a pertussis toxin-sensitive G protein. May have a role in induction of parturition. Possible involvement in cell proliferation and adipose tissue development. Tumor cell motility-stimulating factor.
  • Tissue specificityPredominantly expressed in brain, placenta, ovary, and small intestine. Expressed in a number of carcinomas such as hepatocellular and prostate carcinoma, neuroblastoma and non-small-cell lung cancer. Expressed in body fluids such as plasma, cerebral spinal fluid (CSF), saliva, follicular and amniotic fluids. Not detected in leukocytes. Isoform 1 is more highly expressed in peripheral tissues than in the central nervous system (CNS). Adipocytes only express isoform 1. Isoform 3 is more highly expressed in the brain than in peripheral tissues.
  • Sequence similaritiesBelongs to the nucleotide pyrophosphatase/phosphodiesterase family.
    Contains 2 SMB (somatomedin-B) domains.
  • Post-translational
    modifications
    N-glycosylation, but not furin-cleavage, plays a critical role on secretion and on lysoPLD activity.
  • Cellular localizationSecreted. Secreted by most body fluids including serum and CSF. Also by adipocytes and numerous cancer cells.
  • Information by UniProt
  • Database links
  • Alternative names
    • ATX antibody
    • ATX X antibody
    • Autotaxin antibody
    • Autotaxin t antibody
    • E NPP 2 antibody
    • E-NPP 2 antibody
    • Ectonucleotide pyrophosphatase/phosphodiesterase 2 antibody
    • Ectonucleotide pyrophosphatase/phosphodiesterase family member 2 antibody
    • Enpp2 antibody
    • ENPP2_HUMAN antibody
    • Extracellular lysophospholipase D antibody
    • FLJ26803 antibody
    • LysoPLD antibody
    • NPP2 antibody
    • PD IALPHA antibody
    • PDNP2 antibody
    • Phosphodiesterase I alpha antibody
    • Phosphodiesterase I/nucleotide pyrophosphatase 2 antibody
    • Plasma lysophospholipase D antibody
    see all

Anti-ENPP2 antibody [5H3] images

  • All lanes : Anti-ENPP2 antibody [5H3] (ab77104) at 10 µg/ml

    Lane 1 : Human placenta tissue lysate - total protein (ab29745)
    Lane 2 : Human ovary tissue lysate - total protein (ab30222)
    Lane 3 : Human small intestine tissue lysate - total protein (ab29276)

    Lysates/proteins at 20 µg per lane.

    Secondary
    Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 99 kDa
    Observed band size : 110 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 300 kDa,40 kDa,55 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 20 minutes
  • All lanes : Anti-ENPP2 antibody [5H3] (ab77104) at 10 µg/ml

    Lane 1 : Human kidney tissue lysate - total protein (ab30203)
    Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 99 kDa
    Observed band size : 100 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 180 kDa,250 kDa,50 kDa,65 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 20 minutes
  • IHC image of ENPP2 staining in Human Tonsil FFPE section, performed on a BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab77104, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX
  • ICC/IF image of ab77104 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab77104, 10µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References for Anti-ENPP2 antibody [5H3] (ab77104)

This product has been referenced in:
  • Su SC  et al. Autotaxin-lysophosphatidic acid signaling axis mediates tumorigenesis and development of acquired resistance to sunitinib in renal cell carcinoma. Clin Cancer Res 19:6461-72 (2013). Human . Read more (PubMed: 24122794) »

See 1 Publication for this product

Product Wall


Based on the immunogen for ab77104, the antibody should recognize all isoforms, but we have not specifically tested this. The immunogen is the full length ENPP2 protein.

Thank you for contacting Abcam.

I've compiled a list of primary antibodies that have been tested in mouse tissue via western blot. The only ovary-specific antibodies that I found were anti-human (with only 50% homology with the respective mous...

Read More

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this...

Read More

Thank you for contacting us about this issue. Your protocol appears to be correct, and you should be seeing a signal with 50ug per lane and a 1/400 dilution. Sometimes switching from milk to BSA for blocking and diluting the primary will help develop t...

Read More

Thank you for contacting us.
We recommend to run the gel in denaturing and reducing conditions for ab77104. Therefore, SDS PAGE gel should be used. Attached is our WB protocol and you could use it as a reference.
We use HEK 293 (huamn embryon...

Read More

Thank you for your enquiry.

I am sorry to confirm that these antibodies have not been tested in sandwich ELISA (sELISA). They have been tested in indirect ELISA only and I can recommend considering using them in an indirect ELISA rather than...

Read More

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has ca...

Read More

Thank you for taking the time to complete our questionnaire and provide these details. The details you have kindly provided will provide us with vital information for our monitoring of product quality I appreciate the time you have spent in the lab...

Read More

Thank you for contacting us. Ab82590 and ab77104 are both Anti-ENPP2 mouse monoclonals, however,  ab82590 is from the clone [1F8] and ab77104 is from [5H3]. We are sorry if any confusion has been caused in this matter. We have no plans to discontinue e...

Read More

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"